4-arylamino-2-(6-indolylamino)pyrimidine compounds as antibacterial agents

ABSTRACT

The 4-arylamino-2-(6-indolylamino)pyrimidine derivative compounds are antibacterial agents having broad spectrum antibacterial activity. The present 4-arylamino-2-(6-indolylamino)pyrimidine compounds have antimicrobial activity against various susceptible and resistant gram-positive and gram-negative bacteria as well as drug resistant bacteria, such as MRSA and VRSA. The molecular target of these compounds was identified as DNA Gyrase B. Based on their pharmacological profiles, the present compounds may find important clinical applications for severe infectious diseases and tuberculosis.

BACKGROUND 1. Field

The disclosure of the present patent application relates to4-arylamino-2-(6-indolylamino)pyrimidine compounds, and particularly tonovel 4-arylamino-2-(6-indolylamino)pyrimidine compounds asantibacterial agents.

2. Description of the Related Art

Infectious diseases caused by bacterial resistance to antibioticsconstitute an increasing threat to humans on a global scale. Anincreasing number of infectious bacteria, including tuberculosis,pneumonia, salmonellosis, and gonorrhea, are becoming progressivelychallenging to cure owing to the ineffectiveness of currently usedclinical antibiotics and present a serious health threat worldwide inthe medical community. The major concern of this global health threat isthe ability of the microorganisms to develop one or several mechanismsof resistance to antibiotics, rendering them inefficient for therapeutictreatment. The quest for discovering novel scaffolds with antimicrobialproperties is particularly challenging in hospital and healthcaresettings.

Similarly, the emergence of drug resistant bacteria calls for constantdevelopment of new antibacterial agents with the aim of generatingmedicaments that are potent against drug sensitive and resistantbacteria and well tolerated.

DNA gyrase and topoisomerase IV are attractive targets for developingantibacterial agents in order to combat the emerging bacterialresistance. Novobiocin, an aminocoumarin class, has been the onlyATP-competitive inhibitor of these two enzymes used clinically but waswithdrawn recently for toxicity concerns. The steady demand ofgenerating novel DNA Gyrase B/topoisomerase IV inhibitors is paramountimportance for infectious diseases treatment. New compoundsdemonstrating a broad spectrum of activity against infectious bacteriaand diseases including gram positive and gram negative bacteria,tuberculosis mycobacteria, pneumonia, salmonellosis and gonorrhea amongothers, are desired, as well as such compounds demonstrating potentactivity against drug resistant bacteria such as MRSA, VRSA.

A chosen molecular target of new compound derivatives was identified asDNA Gyrase B/topoisomerase IV. Based on their pharmacological profiles,new compounds may find important clinical applications for severeinfectious diseases including tuberculosis, among others.

Tuberculosis (TB) is one of the top 10 leading causes of mortalityworldwide and is the leading cause of death from infectious diseaseamong adults. It is a communicable disease caused by the bacteriumMycobacterium tuberculosis (MTB) that primarily affects the lungs,resulting in pulmonary TB. TB can also infect other sites of the body,causing extrapulmonary TB.

To date, no major changes in the treatment of TB have been made, and thecurrent standard treatment still involves a combination of fourantibiotics (isoniazid, rifampin, pyrazinamide, and ethambutol) givenfor two months followed by isoniazid and rifampicin for an additionalfour months. This anti-TB regimen has been successful in the treatmentof MTB H37Rv. However, the emergence of multidrug-resistant TB (MDR-TB)and extensively drug-resistant TB (XDR-TB), as well as HIV/TBco-infection cases, have made TB control more difficult. Moreover,treating resistant TB can take up to 24 months and might be associatedwith side effects and a low chance of cure. This, in turn, can lead topoor patient compliance, which can also contribute to the development ofresistance.

Despite the efforts to discover new anti-TB compounds, current therapiesare still facing the development of resistance and poor compliance dueto long treatment duration. Therefore, it is evident that there is anurgent need for the development of new potential antimicrobial compoundsgenerally, and anti-TB compounds specifically, that can act on newmolecular targets to overcome drug-resistant MTB strains and to controlthe wide spread of TB.

Thus, new compounds as antibacterial agents solving the aforementionedproblems is desired.

SUMMARY

The present subject matter relates to new4-arylamino-2-(6-indolylamino)pyrimidine compounds as antibacterialagents that have antimicrobial properties against various susceptibleand resistant gram-positive and gram-negative bacteria as well as drugresistant bacteria. Also presented is a production synthesis method forsaid compounds, a pharmaceutical composition comprising the compound asan active ingredient, and methods of treatment using the presentcompounds. The compounds possess enzymatic activity and inhibitionagainst DNA gyrase as well as having antibacterial properties.

In one embodiment, the present subject matter relates to a compoundhaving the formula I:

or a pharmaceutically acceptable salt, ester stereoisomer, or solvatethereof, wherein:

-   -   each of X₁, X₂, and X₃ is independently CH or nitrogen;    -   each of Y₁, Y₂, Y₃, Y₄, and Y₅ is independently CH or nitrogen;    -   R₁ is a C₁-C₆ alkyl or C₃-C₈ cycloalkyl;    -   R₂ is H, C₁-C₆ alkyl, or halogen;    -   R₃, R₄, R₅, R₆, and R₇ may independently be the same or        different and are each selected from the group consisting of H,        C₁-C₆ alkyl, a carboxylic acid, acyl, sulfonyl, amino, C₁-C₆        alkylamino, hydroxyl, C₁-C₆ polyhaloalkyl, C₁-C₆ polyhaloalkoxy,        C₁-C₆ alkoxy, —CONR′R″, —CH₂CO₂R′, —CONHSO₂NR′R″, —SO₂NR′R″, and        —CH₂NR′R″; and    -   R′ and R″ may each independently be selected from the group        consisting of H, C₃-C₈ cycloalkyl, and C₁-C₆ alkyl.

In another embodiment, the present subject matter relates to a compoundhaving the formula I:

or a pharmaceutically acceptable salt, ester stereoisomer, or solvatethereof, wherein:

-   -   X₁ and X₂ are nitrogen;    -   X₃ is CH;    -   Y₁, Y₂, Y₃, and Y₅ are hydrogen;    -   Y₄ is hydrogen or nitrogen;    -   R₁ is methyl;    -   R₂ is hydrogen; and    -   R₂, R₃, R₄, R₅, and R₆ are independently selected from the group        consisting of hydrogen, a carboxylic acid, trifluoromethyl,        trifluoromethoxy, —SO₂NH₂, a methylamino group substituted with        a methyl and a cyclohexyl group, a cyclopropyl group, or a        pentyl group, hydroxy, and —CONHSO₂N(CH₃)₂.

In a further embodiment, the present subject matter relates to acompound selected from the group consisting of:4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(1),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(2),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(3),N—(N,N-dimethylsulfamoyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzamide(4), 5-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)picolinicacid (5),3-((cyclohexyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (6),3-((cyclopropyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (7),3-((methyl(pentyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (8),2-hydroxy-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (9),3-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(10),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(11),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(12), and a pharmaceutically acceptable salt, ester, stereoisomer, orsolvate thereof.

Further contemplated herein are pharmaceutical compositions containingthese compounds, as well as methods of treating various bacterialinfections by administering the present compounds to a patient in needthereof.

These and other features of the present subject matter will becomereadily apparent upon further review of the following specification.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following definitions are provided for the purpose of understandingthe present subject matter and for construing the appended patentclaims.

Definitions

Throughout the application, where compositions are described as having,including, or comprising specific components, or where processes aredescribed as having, including, or comprising specific process steps, itis contemplated that compositions of the present teachings can alsoconsist essentially of, or consist of, the recited components, and thatthe processes of the present teachings can also consist essentially of,or consist of, the recited process steps.

It is noted that, as used in this specification and the appended claims,the singular forms “a”, “an”, and “the” include plural references unlessthe context clearly dictates otherwise.

In the application, where an element or component is said to be includedin and/or selected from a list of recited elements or components, itshould be understood that the element or component can be any one of therecited elements or components, or the element or component can beselected from a group consisting of two or more of the recited elementsor components. Further, it should be understood that elements and/orfeatures of a composition or a method described herein can be combinedin a variety of ways without departing from the spirit and scope of thepresent teachings, whether explicit or implicit herein.

The use of the terms “include,” “includes”, “including,” “have,” “has,”or “having” should be generally understood as open-ended andnon-limiting unless specifically stated otherwise.

The use of the singular herein includes the plural (and vice versa)unless specifically stated otherwise. In addition, where the use of theterm “about” is before a quantitative value, the present teachings alsoinclude the specific quantitative value itself, unless specificallystated otherwise. As used herein, the term “about” refers to a ±10%variation from the nominal value unless otherwise indicated or inferred.

As used herein, “halo” or “halogen” refers to fluoro, chloro, bromo, andiodo.

As used herein, “alkyl” refers to a straight-chain or branched saturatedhydrocarbon group. Examples of alkyl groups include methyl (Me), ethyl(Et), propyl (e.g., n-propyl and z′-propyl), butyl (e.g., n-butyl,z′-butyl, sec-butyl, tert-butyl), pentyl groups (e.g., n-pentyl,z′-pentyl, -pentyl), hexyl groups, and the like. In various embodiments,an alkyl group can have 1 to 40 carbon atoms (i.e., C₁-C₄₀ alkyl group),for example, 1-30 carbon atoms (i.e., C₁-C₃₀ alkyl group). In someembodiments, an alkyl group can have 1 to 6 carbon atoms, and can bereferred to as a “lower alkyl group” or a “C₁-C₆ alkyl group”. Examplesof lower alkyl groups include methyl, ethyl, propyl (e.g., n-propyl andz′-propyl), and butyl groups (e.g., n-butyl, z′-butyl, sec-butyl,tert-butyl). In some embodiments, alkyl groups can be substituted asdescribed herein. An alkyl group is generally not substituted withanother alkyl group, an alkenyl group, or an alkynyl group.

As used herein, “alkenyl” refers to a straight-chain or branched alkylgroup having one or more carbon-carbon double bonds. Examples of alkenylgroups include ethenyl, propenyl, butenyl, pentenyl, hexenyl,butadienyl, pentadienyl, hexadienyl groups, and the like. The one ormore carbon-carbon double bonds can be internal (such as in 2-butene) orterminal (such as in 1-butene). In various embodiments, an alkenyl groupcan have 2 to 40 carbon atoms (i.e., C₂-C₄₀ alkenyl group), for example,2 to 20 carbon atoms (i.e., C₂-C₂₀ alkenyl group) or 2 to 6 carbon atoms(i.e., C₂-C₆ alkenyl group). In some embodiments, alkenyl groups can besubstituted as described herein. An alkenyl group is generally notsubstituted with another alkenyl group, an alkyl group, or an alkynylgroup.

The term “substituted alkyl” as used herein refers to an alkyl group inwhich 1 or more (up to about 5, for example about 3) hydrogen atoms isreplaced by a substituent independently selected from the group: —O, —S,acyl, acyloxy, optionally substituted alkoxy, optionally substitutedamino (wherein the amino group may be a cyclic amine), azido, carboxyl,(optionally substituted alkoxy)carbonyl, amido, cyano, optionallysubstituted cycloalkyl, optionally substituted cycloalkenyl, halogen,hydroxyl, nitro, sulfamoyl, sulfanyl, sulfinyl, sulfonyl, and sulfonicacid. Some of the optional substituents for alkyl are hydroxy, halogenexemplified by chloro and bromo, acyl exemplified by methylcarbonyl;alkoxy, and heterocyclyl exemplified by morpholino and piperidino. Otheralkyl substituents as described herein may further be contemplated.

The term “substituted alkenyl” refers to an alkenyl group in which 1 ormore (up to about 5, for example about 3) hydrogen atoms is replaced bya substituent independently selected from those listed above withrespect to a substituted alkyl. Other alkenyl substituents as describedherein may further be contemplated.

As used herein, “heteroatom” refers to an atom of any element other thancarbon or hydrogen and includes, for example, nitrogen, oxygen, silicon,sulfur, phosphorus, and selenium.

As used herein, “aryl” refers to an aromatic monocyclic hydrocarbon ringsystem or a polycyclic ring system in which two or more aromatichydrocarbon rings are fused (i.e., having a bond in common with)together or at least one aromatic monocyclic hydrocarbon ring is fusedto one or more cycloalkyl and/or cycloheteroalkyl rings. An aryl groupcan have 6 to 24 carbon atoms in its ring system (e.g., C₆-C₂₄ arylgroup), which can include multiple fused rings. In some embodiments, apolycyclic aryl group can have 8 to 24 carbon atoms. Any suitable ringposition of the aryl group can be covalently linked to the definedchemical structure. Examples of aryl groups having only aromaticcarbocyclic ring(s) include phenyl, 1-naphthyl (bicyclic), 2-naphthyl(bicyclic), anthracenyl (tricyclic), phenanthrenyl (tricyclic),pentacenyl (pentacyclic), and like groups. Examples of polycyclic ringsystems in which at least one aromatic carbocyclic ring is fused to oneor more cycloalkyl and/or cycloheteroalkyl rings include, among others,benzo derivatives of cyclopentane (i.e., an indanyl group, which is a5,6-bicyclic cycloalkyl/aromatic ring system), cyclohexane (i.e., atetrahydronaphthyl group, which is a 6,6-bicyclic cycloalkyl/aromaticring system), imidazoline (i.e., a benzimidazolinyl group, which is a5,6-bicyclic cycloheteroalkyl/aromatic ring system), and pyran (i.e., achromenyl group, which is a 6,6-bicyclic cycloheteroalkyl/aromatic ringsystem). Other examples of aryl groups include benzodioxanyl,benzodioxolyl, chromanyl, indolinyl groups, and the like. In someembodiments, aryl groups can be substituted as described herein. In someembodiments, an aryl group can have one or more halogen substituents,and can be referred to as a “haloaryl” group. Perhaloaryl groups, i.e.,aryl groups where all of the hydrogen atoms are replaced with halogenatoms (e.g., —C₆F₅), are included within the definition of “haloaryl”.In certain embodiments, an aryl group is substituted with another arylgroup and can be referred to as a biaryl group. Each of the aryl groupsin the biaryl group can be substituted as disclosed herein.

As used herein, “heteroaryl” refers to an aromatic monocyclic ringsystem containing at least one ring heteroatom selected from oxygen (O),nitrogen (N), sulfur (S), silicon (Si), and selenium (Se) or apolycyclic ring system where at least one of the rings present in thering system is aromatic and contains at least one ring heteroatom.Polycyclic heteroaryl groups include those having two or more heteroarylrings fused together, as well as those having at least one monocyclicheteroaryl ring fused to one or more aromatic carbocyclic rings,non-aromatic carbocyclic rings, and/or non-aromatic cycloheteroalkylrings. A heteroaryl group, as a whole, can have, for example, 5 to 24ring atoms and contain 1-5 ring heteroatoms (i.e., 5-20 memberedheteroaryl group). The heteroaryl group can be attached to the definedchemical structure at any heteroatom or carbon atom that results in astable structure. Generally, heteroaryl rings do not contain O—O, S—S,or S—O bonds. However, one or more N or S atoms in a heteroaryl groupcan be oxidized (e.g., pyridine N-oxide thiophene S-oxide, thiopheneS,S-dioxide). Examples of heteroaryl groups include, for example, the 5-or 6-membered monocyclic and 5-6 bicyclic ring systems shown below:where T is O, S, NH, N-alkyl, N-aryl, N-(arylalkyl) (e.g., N-benzyl),SiH₂, SiH(alkyl), Si(alkyl)₂, SiH(arylalkyl), Si(arylalkyl)₂, orSi(alkyl)(arylalkyl). Examples of such heteroaryl rings includepyrrolyl, furyl, thienyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl,triazolyl, tetrazolyl, pyrazolyl, imidazolyl, isothiazolyl, thiazolyl,thiadiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, indolyl, isoindolyl,benzofuryl, benzothienyl, quinolyl, 2-methylquinolyl, isoquinolyl,quinoxalyl, quinazolyl, benzotriazolyl, benzimidazolyl, benzothiazolyl,benzisothiazolyl, benzisoxazolyl, benzoxadiazolyl, benzoxazolyl,cinnolinyl, 1H-indazolyl, 2H-indazolyl, indolizinyl, isobenzofuyl,naphthyridinyl, phthalazinyl, pteridinyl, purinyl, oxazolopyridinyl,thiazolopyridinyl, imidazopyridinyl, furopyridinyl, thienopyridinyl,pyridopyrimidinyl, pyridopyrazinyl, pyridopyridazinyl, thienothiazolyl,thienoxazolyl, thienoimidazolyl groups, and the like. Further examplesof heteroaryl groups include 4,5,6,7-tetrahydroindolyl,tetrahydroquinolinyl, benzothienopyridinyl, benzofuropyridinyl groups,and the like. In some embodiments, heteroaryl groups can be substitutedas described herein.

The term “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur, and that thedescription includes instances where said event or circumstance occursand instances in which it does not. For example, “optionally substitutedalkyl” means either “alkyl” or “substituted alkyl,” as defined herein.

It will be understood by those skilled in the art with respect to anychemical group containing one or more substituents that such groups arenot intended to introduce any substitution or substitution patterns thatare sterically impractical and/or physically non-feasible.

The term “isomers” or “stereoisomers” as used herein relates tocompounds that have identical molecular formulae but that differ in thearrangement of their atoms in space. Stereoisomers that are not mirrorimages of one another are termed “diastereoisomers” and stereoisomersthat are non-superimposable mirror images are termed “enantiomers,” orsometimes optical isomers. A carbon atom bonded to four non-identicalsubstituents is termed a “chiral center.” Certain compounds herein haveone or more chiral centers and therefore may exist as either individualstereoisomers or as a mixture of stereoisomers. Configurations ofstereoisomers that owe their existence to hindered rotation about doublebonds are differentiated by their prefixes cis and trans (or Z and E),which indicate that the groups are on the same side (cis or Z) or onopposite sides (trans or E) of the double bond in the molecule accordingto the Cahn-Ingold-Prelog rules. All possible stereoisomers arecontemplated herein as individual stereoisomers or as a mixture ofstereoisomers.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the presently described subject matter pertains.

Where a range of values is provided, for example, concentration ranges,percentage ranges, or ratio ranges, it is understood that eachintervening value, to the tenth of the unit of the lower limit, unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the described subject matter. Theupper and lower limits of these smaller ranges may independently beincluded in the smaller ranges, and such embodiments are alsoencompassed within the described subject matter, subject to anyspecifically excluded limit in the stated range. Where the stated rangeincludes one or both of the limits, ranges excluding either or both ofthose included limits are also included in the described subject matter.

Throughout the application, descriptions of various embodiments use“comprising” language. However, it will be understood by one of skill inthe art, that in some specific instances, an embodiment canalternatively be described using the language “consisting essentiallyof” or “consisting of”.

“Subject” as used herein refers to any animal classified as a mammal,including humans, domestic and farm animals, and zoo, sports, and petcompanion animals such as household pets and other domesticated animalssuch as, but not limited to, cattle, sheep, ferrets, swine, horses,poultry, rabbits, goats, dogs, cats and the like.

“Patient” as used herein refers to a subject in need of treatment of acondition, disorder, or disease, such as an acute or chronic airwaydisorder or disease.

For purposes of better understanding the present teachings and in no waylimiting the scope of the teachings, unless otherwise indicated, allnumbers expressing quantities, percentages or proportions, and othernumerical values used in the specification and claims, are to beunderstood as being modified in all instances by the term “about”.Accordingly, unless indicated to the contrary, the numerical parametersset forth in the following specification and attached claims areapproximations that may vary depending upon the desired propertiessought to be obtained. At the very least, each numerical parametershould at least be construed in light of the number of reportedsignificant digits and by applying ordinary rounding techniques.

4-arylamino-2-(6-indolylamino)pyrimidine derivative compounds

In one embodiment, the present subject matter relates to a compoundhaving the formula I:

or a pharmaceutically acceptable salt, ester stereoisomer, or solvatethereof, wherein:

-   -   each of X₁, X₂, and X₃ is independently CH or nitrogen;    -   each of Y₁, Y₂, Y₃, Y₄, and Y₅ is independently CH or nitrogen;    -   R₁ is a C₁-C₆ alkyl or C₃-C₈ cycloalkyl;    -   R₂ is H, C₁-C₆ alkyl, or halogen;    -   R₃, R₄, R₅, R₆, and R₇ may independently be the same or        different and are each selected from the group consisting of H,        C₁-C₆ alkyl, a carboxylic acid, acyl, sulfonyl, amino, C₁-C₆        alkylamino, hydroxyl, C₁-C₆ polyhaloalkyl, C₁-C₆ polyhaloalkoxy,        C₁-C₆ alkoxy, —CONR′R″, —CH₂CO₂R′, —CONHSO₂NR′R″, —SO₂NR′R″, and        —CH₂NR′R″; and R′ and R″ may each independently be selected from        the group consisting of H, C₃-C₈ cycloalkyl, and C₁-C₆ alkyl.

In another embodiment, the present subject matter relates to a compoundof formula I, wherein R₁ is methyl.

In a further embodiment, the present subject matter relates to acompound of formula I, wherein R₂ is hydrogen.

In a still further embodiment, the present subject matter relates to acompound of formula I, wherein each of R₂, R₃, R₄, R₅, and R₆ areindependently selected from the group consisting of hydrogen, acarboxylic acid, trifluoromethyl, trifluoromethoxy, —SO₂NH₂, amethylamino group substituted with a methyl and a cyclohexyl group, acyclopropyl group, or a pentyl group, hydroxy, and —CONHSO₂N(CH₃)₂.

In yet another embodiment, the present subject matter relates to acompound of formula I, wherein each of R₂, R₃, R₅, and R₆ are H and R₄is selected from the group consisting of a carboxylic acid,trifluoromethyl, trifluoromethoxy, and —CONHSO₂N(CH₃)₂.

In still yet another embodiment, the present subject matter relates to acompound of formula I, wherein each of R₂, R₃, and R₅ are H, R₄ is acarboxylic acid, and R₆ is a methylamino group substituted with a methyland a cyclohexyl group, a cyclopropyl group, or a pentyl group.

In an additional embodiment, the present subject matter relates to acompound of formula I, wherein each of R₂, R₃, R₄, and R₆ are H and R₅is selected from the group consisting of a carboxylic acid,trifluoromethyl, and trifluoromethoxy.

In a further embodiment, the present subject matter relates to acompound of formula I, wherein X₁ and X₂ are nitrogen, X₃ is CH, Y₁, Y₂,Y₃, and Y₅ are hydrogen, and Y₄ is hydrogen or nitrogen.

In another embodiment, the present subject matter relates to a compoundof formula I, wherein the compound is selected from the group consistingof: 4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (1),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(2),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(3),N—(N,N-dimethylsulfamoyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzamide(4), 5-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)picolinicacid (5),3-((cyclohexyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (6),3-((cyclopropyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (7),3-((methyl(pentyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (8),2-hydroxy-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (9),3-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(10),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(11),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(12), and a pharmaceutically acceptable salt, ester, stereoisomer, orsolvate thereof.

Said differently, the present subject matter can relate to compounds offormula I selected from the group consisting of:

and a pharmaceutically acceptable salt, ester, stereoisomer, or solvatethereof.

It is to be understood that the present subject matter covers allcombinations of substituent groups referred to herein.

In further embodiments, the 4-arylamino-2-(6-indolylamino)pyrimidinederivative compounds are shown to have promising antimicrobial activityagainst various susceptible and resistant gram-positive andgram-negative bacteria, and to possess enzymatic activity against DNAgyrase and topoisomerase IV enzymes as well as antibacterial properties.

The present compounds may contain, e.g., when isolated in crystallineform, varying amounts of solvents. Accordingly, the present subjectmatter includes all solvates of the present compounds of formula I andpharmaceutically acceptable stereoisomers, esters, and/or salts thereof.Hydrates are one example of such solvates.

Further, the present subject matter includes all mixtures of possiblestereoisomers of the embodied compounds, independent of the ratio,including the racemates.

Salts of the present compounds, or the salts of the stereoisomersthereof, include all inorganic and organic acid addition salts and saltswith bases, especially all pharmaceutically acceptable inorganic andorganic acid addition salts and salts with bases, particularly allpharmaceutically acceptable inorganic and organic acid addition saltsand salts with bases customarily used in pharmacy.

Examples of acid addition salts include, but are not limited to,hydrochlorides, hydrobromides, phosphates, nitrates, sulfates, acetates,trifluoroacetates, citrates, D-gluconates, benzoates,2-(4-hydroxy-benzoyl)benzoates, butyrates, subsalicylates, maleates,laurates, malates, lactates, fumarates, succinates, oxalates, tartrates,stearates, benzenesulfonates (besilates), toluenesulfonates (tosilates),methanesulfonates (mesilates) and 3-hydroxy-2-naphthoates.

Examples of salts with bases include, but are not limited to, lithium,sodium, potassium, calcium, aluminum, magnesium, titanium, ammonium,meglumine and guanidinium salts. The salts include water-insoluble and,particularly, water-soluble salts.

The present compounds, the salts, the stereoisomers and the salts of thestereoisomers thereof may contain, e.g., when isolated in crystallineform, varying amounts of solvents. Included within the present scopeare, therefore, all solvates of the compounds of formula I, as well asthe solvates of the salts, the stereoisomers and the salts of thestereoisomers of the compounds of formula I.

The present compounds may be isolated and purified in a manner known perse, e.g., by distilling off the solvent in vacuo and recrystallizing theresidue obtained from a suitable solvent or subjecting it to one of thecustomary purification methods, such as column chromatography on asuitable support material.

Salts of the compounds of formula I and the stereoisomers thereof can beobtained by dissolving the free compound in a suitable solvent (by wayof non-limiting example, a ketone such as acetone, methylethylketone ormethylisobutylketone; an ether such as diethyl ether, tetrahydrofuraneor dioxane; a chlorinated hydrocarbon such as methylene chloride orchloroform; a low molecular weight aliphatic alcohol such as methanol,ethanol or isopropanol; a low molecular weight aliphatic ester such asethyl acetate or isopropyl acetate; or water) which contains the desiredacid or base, or to which the desired acid or base is then added. Theacid or base can be employed in salt preparation, depending on whether amono- or polybasic acid or base is concerned and depending on which saltis desired, in an equimolar quantitative ratio or one differingtherefrom. The salts are obtained by filtering, reprecipitating,precipitating with a non-solvent for the salt or by evaporating thesolvent. Salts obtained can be converted into the free compounds which,in turn, can be converted into salts. In this manner, pharmaceuticallyunacceptable salts, which can be obtained, for example, as processproducts in the manufacturing on an industrial scale, can be convertedinto pharmaceutically acceptable salts by processes known to the personskilled in the art.

Pure diastereomers and pure enantiomers of the present compounds can beobtained, e.g., by asymmetric synthesis, by using chiral startingcompounds in synthesis and by splitting up enantiomeric anddiastereomeric mixtures obtained in synthesis. Preferably, the purediastereomeric and pure enantiomeric compounds are obtained by usingchiral starting compounds in synthesis.

Enantiomeric and diastereomeric mixtures can be split up into the pureenantiomers and pure diastereomers by methods known to a person skilledin the art. Preferably, diastereomeric mixtures are separated bycrystallization, in particular fractional crystallization, orchromatography. Enantiomeric mixtures can be separated, e.g., by formingdiastereomers with a chiral auxiliary agent, resolving the diastereomersobtained and removing the chiral auxiliary agent. As chiral auxiliaryagents, for example, chiral acids can be used to separate enantiomericbases and chiral bases can be used to separate enantiomeric acids viaformation of diastereomeric salts. Furthermore, diastereomericderivatives such as diastereomeric esters can be formed fromenantiomeric mixtures of alcohols or enantiomeric mixtures of acids,respectively, using chiral acids or chiral alcohols, respectively, aschiral auxiliary agents. Additionally, diastereomeric complexes ordiastereomeric clathrates may be used for separating enantiomericmixtures. Alternatively, enantiomeric mixtures can be split up usingchiral separating columns in chromatography. Another suitable method forthe isolation of enantiomers is enzymatic separation.

Synthesis:

In one embodiment, the present compounds can be prepared according tothe following reaction scheme:

General procedure for the preparation of4-arylamino-2-(6-indolylamino)pyrimidines II:

To a mixture of arylamine (2.0 mmol) and 2,4-dichloropyrimidine I (2.0mmol) in 5 mL ethanol was added N,N-Diisopropylethylamine (10.0 mmol).The mixture was heated at reflux for 18 h. After cooling to roomtemperature, the precipitate formed was filtrated and washed with coldethanol providing the corresponding 4-arylamino-2-chloropyrimidine whichwas engaged in the next step without further purification. To a mixtureof 6-amino-2-methylindole (1.5 mmol) and 4-arylamino-2-chloropyrimidine(1.5 mmol) in 4 mL butanol was added N,N-Diisopropylethylamine (8.0mmol). The mixture was heated at reflux for 18 h. After cooling to roomtemperature, the precipitate formed was filtrated and recrystallizedfrom ethanol providing the title derivatives II.

Pharmaceutical Compositions:

In another embodiment, the present subject matter is directed topharmaceutical compositions comprising a therapeutically effectiveamount of the compounds as described herein together with one or morepharmaceutically acceptable carriers, excipients, or vehicles. In someembodiments, the present compositions can be used for combinationtherapy, where other therapeutic and/or prophylactic ingredients can beincluded therein.

The present subject matter further relates to a pharmaceuticalcomposition, which comprises at least one of the present compoundstogether with at least one pharmaceutically acceptable auxiliary.

In an embodiment, the pharmaceutical composition comprises one or two ofthe present compounds, or one of the present compounds.

Non-limiting examples of suitable excipients, carriers, or vehiclesuseful herein include liquids such as water, saline, glycerol,polyethylene glycol, hyaluronic acid, ethanol, and the like. Suitableexcipients for nonliquid formulations are also known to those of skillin the art. A thorough discussion of pharmaceutically acceptableexcipients and salts useful herein is available in Remington'sPharmaceutical Sciences, 18th Edition. Easton, Pa., Mack PublishingCompany, 1990, the entire contents of which are incorporated byreference herein.

The present compounds are typically administered at a therapeutically orpharmaceutically effective dosage, e.g., a dosage sufficient to providetreatment for an acute or chronic airway disease or disorder.Administration of the compounds or pharmaceutical compositions thereofcan be by any method that delivers the compounds systemically and/orlocally. These methods include oral routes, parenteral routes,intraduodenal routes, and the like.

While human dosage levels have yet to be optimized for the presentcompounds, generally, a daily dose is from about 0.01 to 10.0 mg/kg ofbody weight, for example about 0.1 to 5.0 mg/kg of body weight. Theprecise effective amount will vary from subject to subject and willdepend upon the species, age, the subject's size and health, the natureand extent of the condition being treated, recommendations of thetreating physician, and the therapeutics or combination of therapeuticsselected for administration. The subject may be administered as manydoses as is required to reduce and/or alleviate the signs, symptoms, orcauses of the disease or disorder in question, or bring about any otherdesired alteration of a biological system.

In employing the present compounds for treatment of a bacterialinfection, any pharmaceutically acceptable mode of administration can beused with other pharmaceutically acceptable excipients, including solid,semi-solid, liquid or aerosol dosage forms, such as, for example,tablets, capsules, powders, liquids, suspensions, suppositories,aerosols or the like. The present compounds can also be administered insustained or controlled release dosage forms, including depotinjections, osmotic pumps, pills, transdermal (includingelectrotransport) patches, and the like, for the prolongedadministration of the compound at a predetermined rate, preferably inunit dosage forms suitable for single administration of precise dosages.

The present compounds may also be administered as compositions preparedas foods for foods or animals, including medical foods, functional food,special nutrition foods and dietary supplements. A “medical food” is aproduct prescribed by a physician that is intended for the specificdietary management of a disorder or health condition for whichdistinctive nutritional requirements exist and may include formulationsfed through a feeding tube (referred to as enteral administration orgavage administration).

A “dietary supplement” shall mean a product that is intended tosupplement the human diet and may be provided in the form of a pill,capsule, tablet, or like formulation. By way of non-limiting example, adietary supplement may include one or more of the following dietaryingredients: vitamins, minerals, herbs, botanicals, amino acids, anddietary substances intended to supplement the diet by increasing totaldietary intake, or a concentrate, metabolite, constituent, extract, orcombinations of these ingredients, not intended as a conventional foodor as the sole item of a meal or diet. Dietary supplements may also beincorporated into foodstuffs, such as functional foods designed topromote control of glucose levels. A “functional food” is an ordinaryfood that has one or more components or ingredients incorporated into itto give a specific medical or physiological benefit, other than a purelynutritional effect. “Special nutrition food” means ingredients designedfor a particular diet related to conditions or to support treatment ofnutritional deficiencies.

Generally, depending on the intended mode of administration, thepharmaceutically acceptable composition will contain about 0.1% to 90%,for example about 0.5% to 50%, by weight of a compound or salt of thepresent compounds, the remainder being suitable pharmaceuticalexcipients, carriers, etc.

One manner of administration for the conditions detailed above is oral,using a convenient daily dosage regimen which can be adjusted accordingto the degree of affliction. For such oral administration, apharmaceutically acceptable, non-toxic composition is formed by theincorporation of any of the normally employed excipients, such as, forexample, mannitol, lactose, starch, magnesium stearate, sodiumsaccharine, talcum, cellulose, sodium crosscarmellose, glucose, gelatin,sucrose, magnesium carbonate, and the like. Such compositions take theform of solutions, suspensions, tablets, dispersible tablets, pills,capsules, powders, sustained release formulations and the like.

The present compositions may take the form of a pill or tablet and thusthe composition may contain, along with the active ingredient, a diluentsuch as lactose, sucrose, dicalcium phosphate, or the like; a lubricantsuch as magnesium stearate or the like; and a binder such as starch, gumacacia, polyvinylpyrrolidine, gelatin, cellulose and derivativesthereof, and the like.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc. an active compound as definedabove and optional pharmaceutical adjuvants in a carrier, such as, forexample, water, saline, aqueous dextrose, glycerol, glycols, ethanol,and the like, to thereby form a solution or suspension. If desired, thepharmaceutical composition to be administered may also contain minoramounts of nontoxic auxiliary substances such as wetting agents,emulsifying agents, or solubilizing agents, pH buffering agents and thelike, for example, sodium acetate, sodium citrate, cyclodextrinederivatives, sorbitan monolaurate, triethanolamine acetate,triethanolamine oleate, etc.

For oral administration, a pharmaceutically acceptable non-toxiccomposition may be formed by the incorporation of any normally employedexcipients, such as, for example, pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, talcum, cellulose derivatives,sodium crosscarmellose, glucose, sucrose, magnesium carbonate, sodiumsaccharin, talcum and the like. Such compositions take the form ofsolutions, suspensions, tablets, capsules, powders, sustained releaseformulations and the like.

For a solid dosage form, a solution or suspension in, for example,propylene carbonate, vegetable oils or triglycerides, may beencapsulated in a gelatin capsule. Such diester solutions, and thepreparation and encapsulation thereof, are disclosed in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545, the contents of each of which areincorporated herein by reference. For a liquid dosage form, thesolution, e.g., in a polyethylene glycol, may be diluted with asufficient quantity of a pharmaceutically acceptable liquid carrier,e.g., water, to be easily measured for administration.

Alternatively, liquid or semi-solid oral formulations may be prepared bydissolving or dispersing the active compound or salt in vegetable oils,glycols, triglycerides, propylene glycol esters (e.g., propylenecarbonate) and the like, and encapsulating these solutions orsuspensions in hard or soft gelatin capsule shells.

Other useful formulations include those set forth in U.S. Pat. Nos. Re.28,819 and 4,358,603, the contents of each of which are herebyincorporated by reference.

Another manner of administration is parenteral administration, generallycharacterized by injection, either subcutaneously, intramuscularly orintravenously. Injectables can be prepared in conventional forms, eitheras liquid solutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, glycerol, ethanolor the like. In addition, if desired, the pharmaceutical compositions tobe administered may also contain minor amounts of non-toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agents,solubility enhancers, and the like, such as for example, sodium acetate,sorbitan monolaurate, triethanolamine oleate, cyclodextrins, etc.

Another approach for parenteral administration employs the implantationof a slow-release or sustained-release system, such that a constantlevel of dosage is maintained. The percentage of active compoundcontained in such parenteral compositions is highly dependent on thespecific nature thereof, as well as the activity of the compound and theneeds of the subject. However, percentages of active ingredient of 0.01%to 10% in solution are employable and will be higher if the compositionis a solid which will be subsequently diluted to the above percentages.The composition may comprise 0.2% to 2% of the active agent in solution.Nasal solutions of the active compound alone or in combination withother pharmaceutically acceptable excipients can also be administered.

Formulations of the active compound or a salt may also be administeredto the respiratory tract as an aerosol or solution for a nebulizer, oras a microfine powder for insufflation, alone or in combination with aninert carrier such as lactose. In such a case, the particles of theformulation have diameters of less than 50 microns, for example lessthan 10 microns.

Methods of Use:

The present compounds have valuable pharmaceutical properties, whichmake them commercially utilizable. Accordingly, the present subjectmatter further relates to use of the present compounds for the treatmentof diseases or infections, especially bacterial infections and/ortuberculosis.

In this regard, the present subject matter relates to a method oftreating a bacterial infection in a patient, comprising administering toa patient in need thereof a therapeutically effective amount of acompound as described herein.

In a further embodiment, the bacterial infection treatable herein can becaused by Gram-positive Staphylococcus aureus, Staphylococcus aureusresistant to MRSA, Staphylococcus epidermidis, Bacillus subtilis,Enterococcus faecium, Enterococcus faecium resistant to VRE, orGram-negative organisms including Klebsiella pneumonia, Escherichiacoli, or Pseudomonas aeruginosa, or any combination thereof.

In one specific embodiment, the bacterial infection can be caused byH37Rv type of M. tuberculosis strains. In another specific embodiment,the bacterial infection can be caused by E. coli.

The compounds as described herein are not only new but have veryvaluable antimicrobial properties. These compounds showed a broadspectrum of activity against gram positive and gram-negative bacteria,as well tuberculosis mycobacteria. They also showed potent activityagainst drug resistant bacteria such as MRSA and VRSA. The moleculartarget of these derivatives was identified as DNA Gyrase B. Based ontheir pharmacological profiles, the present compounds may find importantclinical applications for severe infectious diseases and tuberculosis.

The present subject matter also relates to the use of a compound asdescribed herein in the manufacture of a pharmaceutical composition forthe treatment of bacterial infections, such as the diseases or disordersexemplified above. In particular, the present subject matter relates tothe use of a compound as described herein in the manufacture of apharmaceutical composition for the treatment or prophylaxis of bacterialinfection, such as, but not limited to, bacterial infections caused byGram-positive Staphylococcus aureus, Staphylococcus aureus resistant toMRSA, Staphylococcus epidermidis, Bacillus subtilis, Enterococcusfaecium, Enterococcus faecium resistant to VRE, Gram-negative organismsincluding Klebsiella pneumonia, Escherichia coli, or Pseudomonasaeruginosa, H37Rv type of M. tuberculosis strains, E. coli, or anycombination thereof.

The present subject matter further relates to a method of treating orpreventing a disease comprising administering to a patient in needthereof a therapeutically effective amount of at least one of thecompounds herein.

In particular, the present subject matter relates to a method oftreating one of the above-mentioned diseases or disorders comprisingadministering to a patient in need thereof a therapeutically effectiveamount of at least one of the compounds herein.

In an embodiment, the present subject matter relates to a method oftreating a bacterial infection caused by, for example, but not limitedto, Gram-positive Staphylococcus aureus, Staphylococcus aureus resistantto MRSA, Staphylococcus epidermidis, Bacillus subtilis, Enterococcusfaecium, Enterococcus faecium resistant to VRE, Gram-negative organismsincluding Klebsiella pneumonia, Escherichia coli, or Pseudomonasaeruginosa, H37Rv type of M. tuberculosis strains, E. coli, or anycombination thereof comprising administering to a patient in needthereof a therapeutically effective amount of at least one of thecompounds herein.

In the above methods, the patient is preferably a mammal, morepreferably a human.

Furthermore, in the above methods, at least one of the present compoundscan be used. In an embodiment, one or two of the present compounds areused, or one of the present compounds is used.

The following examples relate to various methods of manufacturingcertain specific compounds as described herein.

EXAMPLES Example 14-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(1)

Compound (1) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-aminobenzoic acid as the arylamine.

Elemental Analysis: Calculated C, 66.84; H, 4.77; N, 19.49. Found C,66.75; H, 4.81; N, 19.55.

Example 2N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(2)

Compound (2) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-(trifluoromethyl)aniline as the arylamine.

Elemental Analysis: Calculated C, 62.66; H, 4.21; N, 18.27. Found C,62.60; H, 4.13; N, 18.36.

Example 3N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(3)

Compound (3) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-(trifluoromethoxy)aniline as the arylamine.

Elemental Analysis: Calculated C, 60.15; H, 4.04; N, 17.54. Found C,60.09; H, 4.07; N, 17.49.

Example 4N—(N,N-dimethylsulfamoyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzamide(4)

Compound (4) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-amino-N—(N,N-dimethylsulfamoyl)benzamide as the arylamine.

Elemental Analysis: Calculated C, 56.76; H, 4.98; N, 21.06. Found C,56.82; H, 5.07; N, 21.09.

Example 55-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)picolinic acid(5)

Compound (5) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using5-aminopicolinic acid as the arylamine.

Elemental Analysis: Calculated C, 63.33; H, 4.48; N, 23.32. Found C,63.29; H, 4.53; N, 23.36.

Example 63-((cyclohexyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (6)

Compound (6) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-amino-3-((cyclohexyl(methyl)amino)methyl)benzoic acid as thearylamine.

Elemental Analysis: Calculated C, 62.56; H, 4.75; N, 18.91. Found C,62.51; H, 4.78; N, 18.84.

Example 73-((cyclopropyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (7)

Compound (7) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-amino-3-((cyclopropyl(methyl)amino)methyl)benzoic acid as thearylamine.

Elemental Analysis: Calculated C, 67.86; H, 5.92; N, 18.99. Found C,67.88; H, 5.88; N, 19.03.

Example 83-((methyl(pentyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (8)

Compound (8) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-amino-3-((methyl(pentyl)amino)methyl)benzoic acid as the arylamine.

Elemental Analysis: Calculated C, 68.62; H, 6.83; N, 17.78. Found C,68.57; H, 6.80; N, 17.75.

Example 92-hydroxy-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (9)

Compound (9) was obtained in accordance with the general procedure forthe preparation of 4-arylamino-2-(6-indolylamino)pyrimidines II using4-amino-2-hydroxybenzoic acid as the arylamine.

Elemental Analysis: Calculated C, 63.99; H, 4.56; N, 18.66. Found C,64.03; H, 4.62; N, 18.68.

Example 103-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(10)

Compound (10) was obtained in accordance with the preparation of2,4-diarylaminopyrimidines II using 3-aminobenzoic acid as thearylamine.

Elemental Analysis: Calculated C, 66.84; H, 4.77; N, 19.49. Found C,66.91; H, 4.79; N, 19.56.

Example 11N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(11)

Compound (11) was obtained in accordance with the preparation of2,4-diarylaminopyrimidines II using 3-(trifluoromethoxy)aniline as thearylamine.

Elemental Analysis: Calculated C, 60.15; H, 4.04; N, 17.54. Found C,60.12; H, 4.08; N, 17.49.

Example 12N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(12)

Compound (12) was obtained in accordance with the preparation of2,4-diarylaminopyrimidines II using 3-(trifluoromethyl)aniline as thearylamine.

Elemental Analysis: Calculated C, 62.66; H, 4.21; N, 18.27. Found C,63.58; H, 4.24; N, 18.30.

Example 13 Antibacterial Activity Evaluation

The following example relates to the antibacterial effectiveness ofcertain specific compounds as described herein.

The compounds of examples 1-12 were evaluated for in vitro antibacterialactivity according to the reported procedure (Methods for DilutionAntimicrobial Susceptibility Tests for Bacteria That Grow Aerobically;Approved Standard-Tenth Edition. CLSI document M07-A10. Wayne. PA:Clinical and Laboratory Standards Institute. 2015) against bacterialstrains comprising Gram-positive Staphylococcus aureus, Staphylococcusaureus resistant to MRSA, epidermidis, Bacillus subtilis, Enterococcusfaecium, Enterococcus faecium resistant o VRE, and Gram-negativeorganisms including Klebsiella pneumonia, Escherichia coli, Pseudomonasaeruginosa.

The biological results demonstrated that the aforementioned compoundspossessed favorable antimicrobial activity.

By way of example, the compound of Example 8 displayed promisingantibacterial activity against various drug sensitive and drug resistantbacterial strains as reported in Table 1.

TABLE 1 Antibacterial activity of compound of Example 8 MIC μg/mlMicroorganisms Example 8 Ciprofloxacin Vancomycin Staphylococcus aureus4 0.5 1 ATCC 29213 Staphylococcus aureus 4 16 1 ATCC BAA-1556 (MERSAresistant) Staphylococcus epidermidis 8 0.25 1 Bacillus subtilis MTCC441 2 0.25 0.5 Enterococcus faecium 2 4 0.5 ATCC 35667 Enterococcusfaecium 4 16 >128 ATCC 51559 (VRE resistant) Klebsiella pneumonia 20.75 >128 MTCC 618 Escherichia coli MTCC 443 32 0.05 >128 Escherichiacoli 2 0.05 >128 MTCC 443 + PABN^(a)

Example 14 Anti-Tubercular Properties

The antitubercular property of the compounds of Examples 1-12 wasevaluated towards H37Rv type of M. tuberculosis strains. The inoculumwas prepared from fresh Lowenstein-Jensen re-suspended in 7H9-5 medium(7H9 broth, 0.5% glycerol, 0.1% casitone, supplemented with oleicacid/albumin/dextrose/catalase (OADC), adjusted to a McFarland tube No.1 and further diluted using sterile saline 1:20; 100 μL was used asinoculums. Each drug stock solution was thawed and diluted in 7H9-S atfourfold the final highest concentration assessed. Using one hundredmicroliter volume of 7H9-S broth, serial two-fold dilution was preparedof each drug directly in a sterile 96-well micro-titer plate. On eachmicroplate, a growth control that does not contain antibiotic andcomposed of a sterile control were also effectuated. As water and mediacommonly evaporate from the closest wells to the plate perimeter duringculture, sterile water was added to the perimeters of the wells toprevent evaporation during the incubation. The plate was covered thensealed in plastic bag and incubated at a temperature of 37° C. in normalatmosphere. After seven days of incubation, alamar blue solution (30 mL)was added to each well and the plate was re-incubated overnight. Weadded 50 μL of alamar blue having 0.25 μM concentration. A change incolor from blue (oxidized state) to pink (reduced) indicated the growthof bacteria and MIC was defined as the lowest concentration of drug thatprevented this change in color.

The biological results demonstrated that the compounds of the compoundspossessed favorable anti-mycobacterial activity.

By way of example, the compound of Example 8 displayed a promisinganti-mycobacterial activity of 8 micro-g/mL against Mycobacteriumtuberculosis H37Rv strain.

Example 15

Evaluation of Inhibitory Activities on E. coli DNA Gyrase.

Inhibitory potencies were investigated utilizing Inspiralis assay onstreptavidin-coated 96-well microtiter plates. First, the plates wererehydrated with buffer (20 mM Tris-HCl with pH 7.6, 0.01% w/v BSA, 0.05%v/v Tween 20, 137 mM NaCl) and the biotinylated oligonucleotide was thenimmobilized. After washing off the unbound oligonucleotide, the enzymetest was performed. The reaction volume of 30 μL in buffer (35 mM Tris xHCl with pH 7.5, 4 mM MgCl2, 24 mM KCl, 2 mM DTT, 1.8 mM spermidine, 1mM ATP, 6.5% w/v glycerol, 0.1 mg/mL albumin) contained 1.5 U of DNAgyrase from E. coli, 0.75 μg of relaxed pNO1 plasmid, and 3 μL solutionof the inhibitor in 10% DMSO and 0.008% Tween 20 (Polysorbate 20).Reaction solutions were incubated at 37° C. for 30 min. After that, theTF buffer (50 mM NaOAc with pH 5.0, 50 mM NaCl and 50 mM MgCl2) wasadded to terminate the enzymatic reaction. After additional incubationfor 30 min at rt, during which biotinoligonucleotide-plasmid triplex wasformed, the unbound plasmid was washed off using TF buffer and SybrGOLDin T10 buffer (10 mM Tris HCl with pH 8.0 and 1 mM EDTA) was added. Thefluorescence was measured with a microplate reader (BioTek Synergy H4,excitation: 485 nm, emission: 535 nm). Initial screening was done at 100or 10 M concentration of inhibitors. For the most active inhibitors IC50was determined using seven concentrations of tested compounds. GraphPadPrism software was used to calculate the IC50 values. The result isgiven as the average value of three independent measurements. Novobiocinwas used as a positive control (Discovery of 4,5,6,7Tetrahydrobenzo[1,2-d]thiazoles as Novel DNA Gyrase Inhibitors Targetingthe ATP-Binding Site, J M C, 2015) The biological results demonstratedthat the compounds of the compounds possessed favorable DNA Gyraseactivity.

By way of example, the compound of Example 8 displayed promising E. coliDNA Gyrase of 315 nM. In the same experimental condition, the referencecontrol Novabiocin inhibited E. coli DNA Gyrase activity of 150 nM.

It is to be understood that the 4-arylamino-2-(6-indolylamino)pyrimidinederivative compounds as antibacterial agents are not limited to thespecific embodiments or examples described above, but encompasses anyand all embodiments within the scope of the generic language of thefollowing claims enabled by the embodiments described herein, orotherwise shown in the drawings or described above in terms sufficientto enable one of ordinary skill in the art to make and use the claimedsubject matter.

We claim:
 1. A compound having the formula I:

or a pharmaceutically acceptable salt, ester stereoisomer, or solvatethereof, wherein: each of X₁, X₂, and X₃ is independently CH ornitrogen; each of Y₁, Y₂, Y₃, Y₄, and Y₅ is independently CH ornitrogen; R₁ is a C₁-C₆ alkyl or C₃-C₈ cycloalkyl; R₂ is H, C₁-C₆ alkyl,or halogen; R₃, R₄, R₅, R₆, and R₇ may independently be the same ordifferent and are each selected from the group consisting of H, C₁-C₆alkyl, a carboxylic acid, acyl, sulfonyl, amino, C₁-C₆ alkylamino,hydroxyl, C₁-C₆ polyhaloalkyl, C₁-C₆ polyhaloalkoxy, C₁-C₆ alkoxy,—CONR′R″, —CH₂CO₂R′, —CONHSO₂NR′R″, —SO₂NR′R″, and —CH₂NR′R″; and R′ andR″ may each independently be selected from the group consisting of H,C₃-C₈ cycloalkyl, and C₁-C₆ alkyl.
 2. The compound of claim 1, whereinR₁ is methyl.
 3. The compound of claim 1, wherein R₂ is hydrogen.
 4. Thecompound of claim 1, wherein each of R₂, R₃, R₄, R₅, and R₆ areindependently selected from the group consisting of hydrogen, acarboxylic acid, trifluoromethyl, trifluoromethoxy, —SO₂NH₂, amethylamino group substituted with a methyl and a cyclohexyl group, acyclopropyl group, or a pentyl group, hydroxy, and —CONHSO₂N(CH₃)₂. 5.The compound of claim 1, wherein each of R₂, R₃, R₅, and R₆ are H and R₄is selected from the group consisting of a carboxylic acid,trifluoromethyl, trifluoromethoxy, and —CONHSO₂N(CH₃)₂.
 6. The compoundof claim 1, wherein each of R₂, R₃, and R₅ are H, R₄ is a carboxylicacid, and R₆ is a methylamino group substituted with a methyl and acyclohexyl group, a cyclopropyl group, or a pentyl group.
 7. Thecompound of claim 1, wherein each of R₂, R₃, R₄, and R₆ are H and R₅ isselected from the group consisting of a carboxylic acid,trifluoromethyl, and trifluoromethoxy.
 8. The compound of claim 1,wherein X₁ and X₂ are nitrogen, X₃ is CH, Y₁, Y₂, Y₃, and Y₅ arehydrogen, and Y₄ is hydrogen or nitrogen.
 9. A pharmaceuticallyacceptable composition comprising a therapeutically effective amount ofthe compound of claim 1 and a pharmaceutically acceptable carrier.
 10. Amethod of treating a bacterial infection in a patient, comprisingadministering to a patient in need thereof a therapeutically effectiveamount of the compound of claim
 1. 11. The method of claim 10, whereinthe bacterial infection is caused by Gram-positive Staphylococcusaureus, Staphylococcus aureus resistant to MRSA, Staphylococcusepidermidis, Bacillus subtilis, Enterococcus faecium, Enterococcusfaecium resistant to VRE, or Gram-negative organisms includingKlebsiella pneumonia, Escherichia coli, or Pseudomonas aeruginosa. 12.The method of claim 10, wherein the bacterial infection is caused byH37Rv type of M. tuberculosis strains.
 13. The method of claim 11,wherein the bacterial infection is caused by E. coli.
 14. A compoundhaving the formula I:

or a pharmaceutically acceptable salt, ester stereoisomer, or solvatethereof, wherein: X₁ and X₂ are nitrogen; X₃ is CH; Y₁, Y₂, Y₃, and Y₅are hydrogen; Y₄ is hydrogen or nitrogen; R₁ is methyl; R₂ is hydrogen;and R₂, R₃, R₄, R₅, and R₆ are independently selected from the groupconsisting of hydrogen, a carboxylic acid, trifluoromethyl,trifluoromethoxy, —SO₂NH₂, a methylamino group substituted with a methyland a cyclohexyl group, a cyclopropyl group, or a pentyl group, hydroxy,and —CONHSO₂N(CH₃)₂.
 15. A pharmaceutical composition comprising atherapeutically effective amount of the compound of claim 14 and apharmaceutically acceptable carrier.
 16. A method of treating abacterial infection in a patient, comprising administering to a patientin need thereof a therapeutically effective amount of the compound ofclaim
 14. 17. A compound selected from the group consisting of:4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(1),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(2),N2-(2-methyl-1H-indol-6-yl)-N4-(4-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(3),N—(N,N-dimethylsulfamoyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzamide(4), 5-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)picolinicacid (5),3-((cyclohexyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (6),3-((cyclopropyl(methyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (7),3-((methyl(pentyl)amino)methyl)-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (8),2-hydroxy-4-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoicacid (9),3-((2-((2-methyl-1H-indol-6-yl)amino)pyrimidin-4-yl)amino)benzoic acid(10),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethoxy)phenyl)pyrimidine-2,4-diamine(11),N2-(2-methyl-1H-indol-6-yl)-N4-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine(12), and a pharmaceutically acceptable salt, ester, stereoisomer, orsolvate thereof.
 18. A pharmaceutically acceptable compositioncomprising a therapeutically effective amount of the compound of claim17 and a pharmaceutically acceptable carrier.
 19. A method of treating abacterial infection in a patient, comprising administering to a patientin need thereof a therapeutically effective amount of the compound ofclaim 17.